Expression and Characterization of Mouse Notch1 \u3ci\u3eAbruptex\u3c/i\u3e EGF Repeats in \u3ci\u3eE. coli\u3c/i\u3e

Notch1 is a transmembrane protein involved in cell signaling, which is found to be linked to various genetic and neurodegenerative diseases. The extracellular domain of the Notch1 protein consists of 36 tandem epidermal growth factor-like (EGF) repeats, which are directly involved in ligand binding and are heavily glycosylated. EGF 24-29 repeats lie in the Abruptex region of the Notch1 protein, and regulate the ligand binding activity through structural changes. The role of glycosylation in ligand binding events and the overall structure of the Abruptex region are not known. Also, the role of calcium binding in Notch activation or inhibition is not clear. The goal of this project is to use NMR spectroscopy to study the structure of EGF domains with and without glycosylation, and a three dimensional structure of the Abruptex EGF repeats will be determined. The structures will provide a model to the Abruptex region of Notch1 and will further aid in our understanding of ligand binding activity in the extracellular domain of the Notch1